| Assay Method Information | |
| | hDPP9 Inhibition Assay |
| Description: | Table 4: Compounds were tested in a biochemical inhibition assay using human dipeptidylpeptidase 9 (hDPP9) enzyme (R&D, #5419-SE-010) and the substrate Gly-Pro-amino-methylcoumarin (Enzo #BML-P189-0005) at 10 μM FAC. The enzymatic reactions were conducted in duplicate at rt for 30 min in 50 μL assay buffer (50 mM Tris, pH 7.5, 0.1% BSA). Compound solutions (in DMSO) at 10 CR, 3-fold dilution series were prepared in assay buffer ten-fold higher than the final concentration, and 5 μL of the dilution was added to a 50 μL reaction so that the highest compound concentration was 10 μM FAC and the concentration of DMSO was 1% in all wells. The plates were read on a PHERAstarPLUS (BMG) using kinetic mode with excitation 340 nm and emission 460 nm. Data were analyzed in Graph Pad Prism. IC50 values were determined by plotting % inhibition versus log compound concentration and using a one site dose response model. Raw data signals were normalized using 1% DMSO as 0% control and no enzyme as 100% inhibitor control. |
| Affinity data for this assay | |
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